Thermo Scientific™ Pierce™ EDC, No-Weigh™ Format

Catalog No.	Quantity
PI22980	5 g
PIA35391	10 x 1 mg
PI77149	10 mg
PI22981	25 g

Catalog No. PI22980 Supplier Thermo Scientific™ Supplier No. 22980

Description

Immobilize or conjugate proteins using 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (CAS 25952-53-8), a water-soluble, carboxyl-to-amine crosslinker.

Thermo Scientific™ Pierce EDC is a water-soluble carbodiimide crosslinker that activates carboxyl groups for spontaneous reaction with primary amines, enabling peptide immobilization and hapten-carrier protein conjugation.

EDC is the abbreviation for 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, a crosslinker that has been used in diverse applications such as forming amide bonds in peptide synthesis, attaching haptens to carrier proteins to form immunogens, labeling nucleic acids through 5Æphosphate groups and creating amine-reactive NHS-esters of biomolecules. EDC reacts with a carboxyl to form an amine-reactive O-acylisourea intermediate. If this intermediate does not encounter an amine, it will hydrolyze and regenerate the carboxyl group. In the presence of N-hydroxysulfosuccinimide (Sulfo-NHS, Part No. 24510), EDC can be used to convert carboxyl groups to amine-reactive Sulfo-NHS esters.

Highlights:

• Zero-length crosslinker
• Numerous chemical conjugates can be synthesized via available –COOH and –NH2 groups
• Amide bond formed provides a neutral linkage, which is ideal for preparing peptides and antigens
• Adding Sulfo-NHS (Part No. 24510) enhances the coupling reaction at physiological pH conditions
• Reactive groups: carbodiimide
• Reactive toward: carboxyl and amino groups

Specifications

• Chemical Reactivity: Amine-Carboxyl
• Cleavable: No
• Description: EDC
• Molecular Weight (g/mol): 191.7
• PEGylated: No
• Spacer Arm Length: 0.0 Å
• Content And Storage: Upon receipt store desiccated at -20°C.
• Cell Permeability: No
• Shipping Condition: Wet Ice
• Product Line: Pierce
• Labeling Method: Chemical Labeling
• Crosslinker Type: Heterobifunctional
• Reactive Moiety: Carbodiimide
• Spacer: Short (<10 Å)
• Form: Powder
• Quantity: 5 g
• Solubility: Water
• Format: Standard, Single-use, Premium-grade
• Water Soluble: Yes
• Product Type: Crosslinker

Frequently Asked Questions (FAQs)

Can NHS and EDC stock solutions be made for long-term storage before use?

No. EDC is very unstable in aqueous environments and must be dissolved immediately before use. NHS and Sulfo-NHS are relatively stable in solution but best results are obtained when they are dissolved immediately before use. Store these compounds desiccated at 4°C.

How are NHS and Sulfo-NHS used?

These compounds are used to modify a carboxyl group to form an amine-reactive ester. This is accomplished by mixing NHS with a carboxyl-containing molecule and the carbodiimide EDC (Cat. No. 22980, 22981, 77149, A35391). EDC causes a dehydration reaction between the carboxyl and the NHS hydroxyl group, giving rise to an NHS-ester-activated molecule. The activated molecule can then be reacted spontaneously with a primary amine-containing molecule. Although the carboxyl-molecules can be made to react directly with amines using EDC, the reaction is much more efficient with NHS because a stable intermediate is created. In fact, molecules that are activated as NHS esters can be dried and stored for later reaction to amine-containing targets.

What are NHS and Sulfo-NHS used for?

These compounds are used in conjunction with the crosslinker EDC (Cat. No. 22980, 22981, 77149, A35391) to synthesize amine-reactive labeling reagents, crosslinkers and conjugation compounds. Any compound containing a carboxylic acid (-COOH), such as a protein, or biotin or peptide, can be activated with NHS or Sulfo-NHS to form an NHS ester that will spontaneously react to form covalent amide linkages with proteins and other molecules that contain primary amines (-NH2).

What is the difference between the Thermo Scientific CarboLink (now Glycolink) and CarboxyLink Resins?

The CarboxyLink Resin is for immobilizing carboxyl-containing biomolecules after EDC activation. CarboLink Resin is hydrazide-activated and immobilizes glycoproteins. Carbolink has been replaced with glycolink immobilization kits and resins. Glycolink can also be used to immobilize steroids or other molecules containing ketones. Both resins are capable of coupling ligands via carboxylic acids using EDC (Cat. No. 22980). Both immobilization chemistries are available on UltraLink Resin as UltraLink Hydrazide (glycoproteins) and UltraLink DADPA (carboxyl) Resins.

How do CarboxyLink and DADPA UltraLink Resins immobilize molecules?

These resins are derivatized with diaminodipropylamine (DADPA). DADPA has a terminal primary amine that enables covalent immobilization of peptides or other carboxyl-containing (-COOH) molecules. When incubated with the resin and the carbodiimide crosslinker EDC (included in the CarboxyLink Immobilization Kit), carboxyl-containing molecules become permanently attached to the resin by stable amide bonds. The immobilized molecules can then be used in affinity purification procedures. CarboxyLink/DADPA UltraLink Coupling Resins can also be used to immobilize other kinds of molecules using alternative amine-reactive crosslinking chemistries.

Can you provide the shelf-life for EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride)?

EDC (1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride) is covered under our general 1-year warranty and is guaranteed to be fully functional for 12 months from the date of shipment, if stored as recommended. Please see section 8.1 of our Terms & Conditions of Sale (https://www.thermofisher.com/content/dam/LifeTech/Documents/PDFs/Terms-and-Conditions-of-Sale.pdf) for more details.

Is your EDAC compound the same as your EDC compound?

Yes, both acronyms are used for the same compound. The chemical name for EDC is 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide and the chemical name for EDAC is N-3-(3-dimethylaminopropyl)-N-ethyl-carbodiimide. Both of these compounds are structurally equivalent.

I'm having problems with coupling our molecule onto the gel when using EDC and CarboxyLink Coupling Resin. How does the buffer pH affect the coupling efficiency onto the gel? Is there any other coupling buffer that can be used other than 0.1 M MES buffer?

There are two reactions occurring when a molecule is conjugated to CarboxyLink Resin in the presence of EDC. These reactions may occur simultaneously (at pH 7.0-7.2) or separately (moving from an acidic pH to a slightly alkaline pH). Adding sulfo-NHS to the reaction along with EDC helps increase the yield of the reaction.

The activation reaction of the molecule’s carboxyls with EDC and Sulfo-NHS is most efficient at pH 4.5-7.2; however, the reaction of NHS-activated or sulfo-NHS-activated molecules with primary amines is most efficient at pH 7-8. For best results, perform the first reaction in MES buffer (or other non-amine, non-carboxylate buffer) at pH 4.7-6, then raise the pH to 7.2-7.5 with phosphate buffer (or other non-amine buffer) immediately before reaction to the amine-containing molecule. The reaction should be done with EDC that is freshly prepared. 

Do you offer cross-linkers similar to EDC (Cat. No. 22980) that have flexibility with spacer lengths?

Unfortunately, we do not have a carboxyl-reactive crosslinker that provides a spacer arm to give distance between the two molecules to be linked. We do offer amine/sulfhydryl reactive crosslinkers that are available with various lengths of spacer arms.

What are the general characteristics of N-hydroxysuccinimide esters (NHS Esters)?

NHS esters are reactive groups formed by EDC-activation of carboxylate molecules. NHS ester-activated crosslinkers and labeling compounds react with primary amines in slightly alkaline conditions (pH 7.2-8.5) to yield stable amide bonds. The reaction releases N-hydroxysuccinimide (MW 115), which can be removed easily by dialysis or desalting. Primary amine buffers such as Tris or TBS are not compatible because they compete for reaction; however, in some procedures, it is useful to add Tris or glycine buffer at the end of a conjugation procedure to quench (stop) the reaction.
Sulfo-NHS esters are identical to NHS esters except that they contain a sulfonate (–SO3) group on the N-hydroxysuccinimide ring. This charged group has no effect on the reaction chemistry, but it does tend to increase the water-solubility of crosslinkers containing them. In addition, the charged group prevents sulfo-NHS crosslinkers from permeating cell membranes, enabling them to be used for cell surface crosslinking methods.

What are the general characteristics of carbodiimides?

EDC and other carbodiimides are zero-length crosslinkers; they cause direct conjugation of carboxylates (-COOH) to primary amines (-NH2) without becoming part of the final crosslink (amide bond) between target molecules. EDC crosslinking reactions must be performed in conditions devoid of extraneous carboxyls and amines. Because peptides and proteins contain multiple carboxyls and amines, direct EDC-mediated crosslinking usually causes random polymerization of polypeptides. Nevertheless, this reaction chemistry is used widely in immobilization procedures (e.g., attaching proteins to a carboxylated surface) and in immunogen preparation (e.g., attaching a small peptide to a large carrier protein).

What is the difference between GlycoLink and CarboxyLink Gels?

GlycoLink gel is hydrazide-activated crosslinked beaded agarose, and it is useful for coupling glycoproteins via aldehydes formed from their sugars by sodium meta-periodate oxidation. Reaction of aldehydes with hydrazide- activated resin is catalyzed by aniline resulting in >90% coupling in 4 hours or less. CarboxyLink gel on the other hand is crosslinked beaded agarose activated with diaminodipropylamine (DADPA) and is useful for immobilizing carboxyl- containing biomolecules after EDC activation. Actually, both resins can be used with EDC to couple ligands via carboxylic acids.

Note: Both immobilization chemistries are available on UltraLink Support as UltraLink Hydrazide and UltraLink DADPA respectively.

What is the difference between SulfoLink and CarboxyLink Coupling Resin supports?

SulfoLink Coupling Resin is iodoacetyl-activated agarose that reacts with reduced thiols to form a stable thioether linkage. CarboxyLink Coupling Resin is an amine-derivatized agarose that utilizes the carbodiimide crosslinker, EDC to conjugate the carbon of carboxylic acid to the nitrogen of the immobilized primary amine (hydrazide) to form a permanent amide bond.

What are the benefits of using a CarboxyLink support?

CarboxyLink can be used to immobilize any protein or peptide via the C-terminus or aspartic or glutamic acid residues. Diaminodiproylamine (DADPA) is immobilized onto an agarose support. The carbodiimide crosslinker, EDC facilitates the formation of an amide bond between the carbon of the carboxylic acid and the nitrogen on the terminal amine of DADPA. This support is compatible with 50% organic solvent to improve immobilization of hydrophobic peptides.

For Research Use Only. Not for use in diagnostic procedures.